CN114315784A - Histidine-labeled fluorescent probe and preparation method and application thereof - Google Patents
Histidine-labeled fluorescent probe and preparation method and application thereof Download PDFInfo
- Publication number
- CN114315784A CN114315784A CN202111440062.0A CN202111440062A CN114315784A CN 114315784 A CN114315784 A CN 114315784A CN 202111440062 A CN202111440062 A CN 202111440062A CN 114315784 A CN114315784 A CN 114315784A
- Authority
- CN
- China
- Prior art keywords
- ida
- rho
- histidine
- fluorescent
- fluorescent probe
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 239000007850 fluorescent dye Substances 0.000 title claims abstract description 59
- 238000002360 preparation method Methods 0.000 title abstract description 8
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 title description 6
- PYWVYCXTNDRMGF-UHFFFAOYSA-N rhodamine B Chemical class [Cl-].C=12C=CC(=[N+](CC)CC)C=C2OC2=CC(N(CC)CC)=CC=C2C=1C1=CC=CC=C1C(O)=O PYWVYCXTNDRMGF-UHFFFAOYSA-N 0.000 claims abstract description 16
- 102000037865 fusion proteins Human genes 0.000 claims abstract description 15
- 108020001507 fusion proteins Proteins 0.000 claims abstract description 15
- 238000000034 method Methods 0.000 claims abstract description 12
- 239000013522 chelant Substances 0.000 claims abstract description 8
- 239000012535 impurity Substances 0.000 claims abstract description 4
- XLJKHNWPARRRJB-UHFFFAOYSA-N cobalt(2+) Chemical compound [Co+2] XLJKHNWPARRRJB-UHFFFAOYSA-N 0.000 claims abstract description 3
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 claims description 9
- 229910052751 metal Inorganic materials 0.000 claims description 9
- 239000002184 metal Substances 0.000 claims description 9
- NBZBKCUXIYYUSX-UHFFFAOYSA-N iminodiacetic acid Chemical compound OC(=O)CNCC(O)=O NBZBKCUXIYYUSX-UHFFFAOYSA-N 0.000 claims description 7
- 239000002244 precipitate Substances 0.000 claims description 7
- 229940043267 rhodamine b Drugs 0.000 claims description 7
- 238000001914 filtration Methods 0.000 claims description 6
- 238000002156 mixing Methods 0.000 claims description 6
- NQTADLQHYWFPDB-UHFFFAOYSA-N N-Hydroxysuccinimide Chemical compound ON1C(=O)CCC1=O NQTADLQHYWFPDB-UHFFFAOYSA-N 0.000 claims description 5
- 238000006243 chemical reaction Methods 0.000 claims description 5
- 238000001514 detection method Methods 0.000 claims description 4
- 150000002148 esters Chemical class 0.000 claims description 4
- 239000002904 solvent Substances 0.000 claims description 3
- 239000012295 chemical reaction liquid Substances 0.000 claims description 2
- 238000001704 evaporation Methods 0.000 claims description 2
- 238000003756 stirring Methods 0.000 claims description 2
- 238000011160 research Methods 0.000 abstract description 5
- 230000005284 excitation Effects 0.000 abstract description 4
- 229910021645 metal ion Inorganic materials 0.000 abstract description 2
- 238000012984 biological imaging Methods 0.000 abstract 1
- 229910001429 cobalt ion Inorganic materials 0.000 abstract 1
- 238000001308 synthesis method Methods 0.000 abstract 1
- 102000004169 proteins and genes Human genes 0.000 description 13
- 108090000623 proteins and genes Proteins 0.000 description 13
- 239000000523 sample Substances 0.000 description 11
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 9
- 239000000243 solution Substances 0.000 description 9
- 239000000499 gel Substances 0.000 description 8
- 238000001962 electrophoresis Methods 0.000 description 5
- 238000002372 labelling Methods 0.000 description 5
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 239000003153 chemical reaction reagent Substances 0.000 description 4
- 230000035945 sensitivity Effects 0.000 description 4
- 150000003384 small molecules Chemical class 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 3
- 238000004043 dyeing Methods 0.000 description 3
- 238000000119 electrospray ionisation mass spectrum Methods 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- RAXXELZNTBOGNW-UHFFFAOYSA-N imidazole Natural products C1=CNC=N1 RAXXELZNTBOGNW-UHFFFAOYSA-N 0.000 description 3
- MGFYIUFZLHCRTH-UHFFFAOYSA-N nitrilotriacetic acid Chemical compound OC(=O)CN(CC(O)=O)CC(O)=O MGFYIUFZLHCRTH-UHFFFAOYSA-N 0.000 description 3
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 3
- 238000010186 staining Methods 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 2
- QOSSAOTZNIDXMA-UHFFFAOYSA-N Dicylcohexylcarbodiimide Chemical compound C1CCCCC1N=C=NC1CCCCC1 QOSSAOTZNIDXMA-UHFFFAOYSA-N 0.000 description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- NGBGZCUWFVVJKC-IRXDYDNUSA-N Gly-Tyr-Tyr Chemical compound C([C@H](NC(=O)CN)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)C1=CC=C(O)C=C1 NGBGZCUWFVVJKC-IRXDYDNUSA-N 0.000 description 2
- AZEYWPUCOYXFOE-CYDGBPFRSA-N Ile-Arg-Val Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](C(C)C)C(=O)O)N AZEYWPUCOYXFOE-CYDGBPFRSA-N 0.000 description 2
- SENJXOPIZNYLHU-UHFFFAOYSA-N L-leucyl-L-arginine Natural products CC(C)CC(N)C(=O)NC(C(O)=O)CCCN=C(N)N SENJXOPIZNYLHU-UHFFFAOYSA-N 0.000 description 2
- YBAFDPFAUTYYRW-UHFFFAOYSA-N N-L-alpha-glutamyl-L-leucine Natural products CC(C)CC(C(O)=O)NC(=O)C(N)CCC(O)=O YBAFDPFAUTYYRW-UHFFFAOYSA-N 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- NKLPQNGYXWVELD-UHFFFAOYSA-M coomassie brilliant blue Chemical compound [Na+].C1=CC(OCC)=CC=C1NC1=CC=C(C(=C2C=CC(C=C2)=[N+](CC)CC=2C=C(C=CC=2)S([O-])(=O)=O)C=2C=CC(=CC=2)N(CC)CC=2C=C(C=CC=2)S([O-])(=O)=O)C=C1 NKLPQNGYXWVELD-UHFFFAOYSA-M 0.000 description 2
- 230000008878 coupling Effects 0.000 description 2
- 238000010168 coupling process Methods 0.000 description 2
- 238000005859 coupling reaction Methods 0.000 description 2
- 239000008367 deionised water Substances 0.000 description 2
- 229910021641 deionized water Inorganic materials 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000000799 fluorescence microscopy Methods 0.000 description 2
- 108010078144 glutaminyl-glycine Proteins 0.000 description 2
- 108010050848 glycylleucine Proteins 0.000 description 2
- 108010028295 histidylhistidine Proteins 0.000 description 2
- 108010025306 histidylleucine Proteins 0.000 description 2
- 238000003384 imaging method Methods 0.000 description 2
- 108010000761 leucylarginine Proteins 0.000 description 2
- 238000011068 loading method Methods 0.000 description 2
- 108010064235 lysylglycine Proteins 0.000 description 2
- 108010017391 lysylvaline Proteins 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 238000005160 1H NMR spectroscopy Methods 0.000 description 1
- IICCLYANAQEHCI-UHFFFAOYSA-N 4,5,6,7-tetrachloro-3',6'-dihydroxy-2',4',5',7'-tetraiodospiro[2-benzofuran-3,9'-xanthene]-1-one Chemical compound O1C(=O)C(C(=C(Cl)C(Cl)=C2Cl)Cl)=C2C21C1=CC(I)=C(O)C(I)=C1OC1=C(I)C(O)=C(I)C=C21 IICCLYANAQEHCI-UHFFFAOYSA-N 0.000 description 1
- PXKLCFFSVLKOJM-ACZMJKKPSA-N Ala-Asn-Glu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O PXKLCFFSVLKOJM-ACZMJKKPSA-N 0.000 description 1
- CCDFBRZVTDDJNM-GUBZILKMSA-N Ala-Leu-Glu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O CCDFBRZVTDDJNM-GUBZILKMSA-N 0.000 description 1
- MDNAVFBZPROEHO-DCAQKATOSA-N Ala-Lys-Val Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(O)=O MDNAVFBZPROEHO-DCAQKATOSA-N 0.000 description 1
- MDNAVFBZPROEHO-UHFFFAOYSA-N Ala-Lys-Val Natural products CC(C)C(C(O)=O)NC(=O)C(NC(=O)C(C)N)CCCCN MDNAVFBZPROEHO-UHFFFAOYSA-N 0.000 description 1
- MAZZQZWCCYJQGZ-GUBZILKMSA-N Ala-Pro-Arg Chemical compound [H]N[C@@H](C)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCCNC(N)=N)C(O)=O MAZZQZWCCYJQGZ-GUBZILKMSA-N 0.000 description 1
- SFPRJVVDZNLUTG-OWLDWWDNSA-N Ala-Trp-Thr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H]([C@@H](C)O)C(O)=O SFPRJVVDZNLUTG-OWLDWWDNSA-N 0.000 description 1
- VBFJESQBIWCWRL-DCAQKATOSA-N Arg-Ala-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCCNC(N)=N VBFJESQBIWCWRL-DCAQKATOSA-N 0.000 description 1
- HKRXJBBCQBAGIM-FXQIFTODSA-N Arg-Asp-Ser Chemical compound C(C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CO)C(=O)O)N)CN=C(N)N HKRXJBBCQBAGIM-FXQIFTODSA-N 0.000 description 1
- RKRSYHCNPFGMTA-CIUDSAMLSA-N Arg-Glu-Asn Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O RKRSYHCNPFGMTA-CIUDSAMLSA-N 0.000 description 1
- PNQWAUXQDBIJDY-GUBZILKMSA-N Arg-Glu-Glu Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O PNQWAUXQDBIJDY-GUBZILKMSA-N 0.000 description 1
- NKBQZKVMKJJDLX-SRVKXCTJSA-N Arg-Glu-Leu Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O NKBQZKVMKJJDLX-SRVKXCTJSA-N 0.000 description 1
- UVTGNSWSRSCPLP-UHFFFAOYSA-N Arg-Tyr Natural products NC(CCNC(=N)N)C(=O)NC(Cc1ccc(O)cc1)C(=O)O UVTGNSWSRSCPLP-UHFFFAOYSA-N 0.000 description 1
- FOWOZYAWODIRFZ-JYJNAYRXSA-N Arg-Tyr-Met Chemical compound CSCC[C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)NC(=O)[C@H](CCCN=C(N)N)N FOWOZYAWODIRFZ-JYJNAYRXSA-N 0.000 description 1
- APHUDFFMXFYRKP-CIUDSAMLSA-N Asn-Asn-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CC(=O)N)N APHUDFFMXFYRKP-CIUDSAMLSA-N 0.000 description 1
- KDFQZBWWPYQBEN-ZLUOBGJFSA-N Asp-Ala-Asn Chemical compound C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CC(=O)O)N KDFQZBWWPYQBEN-ZLUOBGJFSA-N 0.000 description 1
- NJIKKGUVGUBICV-ZLUOBGJFSA-N Asp-Ala-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC(O)=O NJIKKGUVGUBICV-ZLUOBGJFSA-N 0.000 description 1
- RXBGWGRSWXOBGK-KKUMJFAQSA-N Asp-Lys-Tyr Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O RXBGWGRSWXOBGK-KKUMJFAQSA-N 0.000 description 1
- KPSHWSWFPUDEGF-FXQIFTODSA-N Asp-Pro-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CC(O)=O KPSHWSWFPUDEGF-FXQIFTODSA-N 0.000 description 1
- AHWRSSLYSGLBGD-CIUDSAMLSA-N Asp-Pro-Glu Chemical compound OC(=O)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(O)=O AHWRSSLYSGLBGD-CIUDSAMLSA-N 0.000 description 1
- NAAAPCLFJPURAM-HJGDQZAQSA-N Asp-Thr-Lys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC(=O)O)N)O NAAAPCLFJPURAM-HJGDQZAQSA-N 0.000 description 1
- 241000534000 Berula erecta Species 0.000 description 1
- XOKGKOQWADCLFQ-GARJFASQSA-N Gln-Arg-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CCC(=O)N)N)C(=O)O XOKGKOQWADCLFQ-GARJFASQSA-N 0.000 description 1
- VNTGPISAOMAXRK-CIUDSAMLSA-N Gln-Pro-Ser Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(O)=O VNTGPISAOMAXRK-CIUDSAMLSA-N 0.000 description 1
- XKPACHRGOWQHFH-IRIUXVKKSA-N Gln-Thr-Tyr Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O XKPACHRGOWQHFH-IRIUXVKKSA-N 0.000 description 1
- HTTSBEBKVNEDFE-AUTRQRHGSA-N Glu-Gln-Val Chemical compound CC(C)[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CCC(=O)O)N HTTSBEBKVNEDFE-AUTRQRHGSA-N 0.000 description 1
- QQLBPVKLJBAXBS-FXQIFTODSA-N Glu-Glu-Asn Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O QQLBPVKLJBAXBS-FXQIFTODSA-N 0.000 description 1
- BUZMZDDKFCSKOT-CIUDSAMLSA-N Glu-Glu-Glu Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O BUZMZDDKFCSKOT-CIUDSAMLSA-N 0.000 description 1
- VOORMNJKNBGYGK-YUMQZZPRSA-N Glu-Gly-Met Chemical compound CSCC[C@@H](C(=O)O)NC(=O)CNC(=O)[C@H](CCC(=O)O)N VOORMNJKNBGYGK-YUMQZZPRSA-N 0.000 description 1
- HPJLZFTUUJKWAJ-JHEQGTHGSA-N Glu-Gly-Thr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(O)=O HPJLZFTUUJKWAJ-JHEQGTHGSA-N 0.000 description 1
- GJBUAAAIZSRCDC-GVXVVHGQSA-N Glu-Leu-Val Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(O)=O GJBUAAAIZSRCDC-GVXVVHGQSA-N 0.000 description 1
- TWYFJOHWGCCRIR-DCAQKATOSA-N Glu-Pro-Arg Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCCNC(N)=N)C(O)=O TWYFJOHWGCCRIR-DCAQKATOSA-N 0.000 description 1
- SWDNPSMMEWRNOH-HJGDQZAQSA-N Glu-Pro-Thr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(O)=O SWDNPSMMEWRNOH-HJGDQZAQSA-N 0.000 description 1
- NNQDRRUXFJYCCJ-NHCYSSNCSA-N Glu-Pro-Val Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C(C)C)C(O)=O NNQDRRUXFJYCCJ-NHCYSSNCSA-N 0.000 description 1
- QXUPRMQJDWJDFR-NRPADANISA-N Glu-Val-Ser Chemical compound CC(C)[C@H](NC(=O)[C@@H](N)CCC(O)=O)C(=O)N[C@@H](CO)C(O)=O QXUPRMQJDWJDFR-NRPADANISA-N 0.000 description 1
- FKJQNJCQTKUBCD-XPUUQOCRSA-N Gly-Ala-His Chemical compound NCC(=O)N[C@@H](C)C(=O)N[C@@H](CC1=CNC=N1)C(=O)O FKJQNJCQTKUBCD-XPUUQOCRSA-N 0.000 description 1
- JBRBACJPBZNFMF-YUMQZZPRSA-N Gly-Ala-Lys Chemical compound NCC(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CCCCN JBRBACJPBZNFMF-YUMQZZPRSA-N 0.000 description 1
- IWAXHBCACVWNHT-BQBZGAKWSA-N Gly-Asp-Arg Chemical compound NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CCCN=C(N)N IWAXHBCACVWNHT-BQBZGAKWSA-N 0.000 description 1
- BYYNJRSNDARRBX-YFKPBYRVSA-N Gly-Gln-Gly Chemical compound NCC(=O)N[C@@H](CCC(N)=O)C(=O)NCC(O)=O BYYNJRSNDARRBX-YFKPBYRVSA-N 0.000 description 1
- MOJKRXIRAZPZLW-WDSKDSINSA-N Gly-Glu-Ala Chemical compound [H]NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(O)=O MOJKRXIRAZPZLW-WDSKDSINSA-N 0.000 description 1
- JUBDONGMHASUCN-IUCAKERBSA-N Gly-Glu-His Chemical compound NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](Cc1cnc[nH]1)C(O)=O JUBDONGMHASUCN-IUCAKERBSA-N 0.000 description 1
- QSVCIFZPGLOZGH-WDSKDSINSA-N Gly-Glu-Ser Chemical compound NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(O)=O QSVCIFZPGLOZGH-WDSKDSINSA-N 0.000 description 1
- DENRBIYENOKSEX-PEXQALLHSA-N Gly-Ile-His Chemical compound NCC(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@H](C(O)=O)CC1=CN=CN1 DENRBIYENOKSEX-PEXQALLHSA-N 0.000 description 1
- UESJMAMHDLEHGM-NHCYSSNCSA-N Gly-Ile-Leu Chemical compound NCC(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(C)C)C(O)=O UESJMAMHDLEHGM-NHCYSSNCSA-N 0.000 description 1
- TWTPDFFBLQEBOE-IUCAKERBSA-N Gly-Leu-Gln Chemical compound [H]NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O TWTPDFFBLQEBOE-IUCAKERBSA-N 0.000 description 1
- UHPAZODVFFYEEL-QWRGUYRKSA-N Gly-Leu-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)CN UHPAZODVFFYEEL-QWRGUYRKSA-N 0.000 description 1
- OCPPBNKYGYSLOE-IUCAKERBSA-N Gly-Pro-Met Chemical compound CSCC[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)CN OCPPBNKYGYSLOE-IUCAKERBSA-N 0.000 description 1
- FFJQHWKSGAWSTJ-BFHQHQDPSA-N Gly-Thr-Ala Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(O)=O FFJQHWKSGAWSTJ-BFHQHQDPSA-N 0.000 description 1
- SBVMXEZQJVUARN-XPUUQOCRSA-N Gly-Val-Ser Chemical compound NCC(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(O)=O SBVMXEZQJVUARN-XPUUQOCRSA-N 0.000 description 1
- JBCLFWXMTIKCCB-UHFFFAOYSA-N H-Gly-Phe-OH Natural products NCC(=O)NC(C(O)=O)CC1=CC=CC=C1 JBCLFWXMTIKCCB-UHFFFAOYSA-N 0.000 description 1
- MDBYBTWRMOAJAY-NHCYSSNCSA-N His-Asn-Val Chemical compound CC(C)[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CC1=CN=CN1)N MDBYBTWRMOAJAY-NHCYSSNCSA-N 0.000 description 1
- FIMNVXRZGUAGBI-AVGNSLFASA-N His-Glu-Leu Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O FIMNVXRZGUAGBI-AVGNSLFASA-N 0.000 description 1
- NQKRILCJYCASDV-QWRGUYRKSA-N His-Gly-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CC1=CN=CN1 NQKRILCJYCASDV-QWRGUYRKSA-N 0.000 description 1
- CTJHHEQNUNIYNN-SRVKXCTJSA-N His-His-Asn Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC(N)=O)C(O)=O CTJHHEQNUNIYNN-SRVKXCTJSA-N 0.000 description 1
- STOOMQFEJUVAKR-KKUMJFAQSA-N His-His-His Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1N=CNC=1)C(=O)N[C@@H](CC=1N=CNC=1)C(O)=O)C1=CNC=N1 STOOMQFEJUVAKR-KKUMJFAQSA-N 0.000 description 1
- 108010093488 His-His-His-His-His-His Proteins 0.000 description 1
- QEYUCKCWTMIERU-SRVKXCTJSA-N His-Lys-Asp Chemical compound C1=C(NC=N1)C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(=O)O)C(=O)O)N QEYUCKCWTMIERU-SRVKXCTJSA-N 0.000 description 1
- XIGFLVCAVQQGNS-IHRRRGAJSA-N His-Pro-His Chemical compound C([C@H](N)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CC=1NC=NC=1)C(O)=O)C1=CN=CN1 XIGFLVCAVQQGNS-IHRRRGAJSA-N 0.000 description 1
- FCPSGEVYIVXPPO-QTKMDUPCSA-N His-Thr-Arg Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O FCPSGEVYIVXPPO-QTKMDUPCSA-N 0.000 description 1
- ZHMZWSFQRUGLEC-JYJNAYRXSA-N His-Tyr-Glu Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCC(O)=O)C(O)=O ZHMZWSFQRUGLEC-JYJNAYRXSA-N 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- DPTBVFUDCPINIP-JURCDPSOSA-N Ile-Ala-Phe Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 DPTBVFUDCPINIP-JURCDPSOSA-N 0.000 description 1
- VZIFYHYNQDIPLI-HJWJTTGWSA-N Ile-Arg-Phe Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)N VZIFYHYNQDIPLI-HJWJTTGWSA-N 0.000 description 1
- UMYZBHKAVTXWIW-GMOBBJLQSA-N Ile-Asp-Arg Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)N UMYZBHKAVTXWIW-GMOBBJLQSA-N 0.000 description 1
- LJKDGRWXYUTRSH-YVNDNENWSA-N Ile-Gln-Gln Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N LJKDGRWXYUTRSH-YVNDNENWSA-N 0.000 description 1
- JDAWAWXGAUZPNJ-ZPFDUUQYSA-N Ile-Glu-Arg Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)N JDAWAWXGAUZPNJ-ZPFDUUQYSA-N 0.000 description 1
- DFJJAVZIHDFOGQ-MNXVOIDGSA-N Ile-Glu-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CCCCN)C(=O)O)N DFJJAVZIHDFOGQ-MNXVOIDGSA-N 0.000 description 1
- KYLIZSDYWQQTFM-PEDHHIEDSA-N Ile-Ile-Arg Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@H](C(O)=O)CCCN=C(N)N KYLIZSDYWQQTFM-PEDHHIEDSA-N 0.000 description 1
- UDBPXJNOEWDBDF-XUXIUFHCSA-N Ile-Lys-Val Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(=O)O)N UDBPXJNOEWDBDF-XUXIUFHCSA-N 0.000 description 1
- NJGXXYLPDMMFJB-XUXIUFHCSA-N Ile-Val-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCCN)C(=O)O)N NJGXXYLPDMMFJB-XUXIUFHCSA-N 0.000 description 1
- APQYGMBHIVXFML-OSUNSFLBSA-N Ile-Val-Thr Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)O)N APQYGMBHIVXFML-OSUNSFLBSA-N 0.000 description 1
- LZDNBBYBDGBADK-UHFFFAOYSA-N L-valyl-L-tryptophan Natural products C1=CC=C2C(CC(NC(=O)C(N)C(C)C)C(O)=O)=CNC2=C1 LZDNBBYBDGBADK-UHFFFAOYSA-N 0.000 description 1
- MJOZZTKJZQFKDK-GUBZILKMSA-N Leu-Ala-Gln Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CCC(N)=O MJOZZTKJZQFKDK-GUBZILKMSA-N 0.000 description 1
- KWTVLKBOQATPHJ-SRVKXCTJSA-N Leu-Ala-Lys Chemical compound C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC(C)C)N KWTVLKBOQATPHJ-SRVKXCTJSA-N 0.000 description 1
- ILJREDZFPHTUIE-GUBZILKMSA-N Leu-Asp-Glu Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O ILJREDZFPHTUIE-GUBZILKMSA-N 0.000 description 1
- VWHGTYCRDRBSFI-ZETCQYMHSA-N Leu-Gly-Gly Chemical compound CC(C)C[C@H](N)C(=O)NCC(=O)NCC(O)=O VWHGTYCRDRBSFI-ZETCQYMHSA-N 0.000 description 1
- HYIFFZAQXPUEAU-QWRGUYRKSA-N Leu-Gly-Leu Chemical compound CC(C)C[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CC(C)C HYIFFZAQXPUEAU-QWRGUYRKSA-N 0.000 description 1
- QNBVTHNJGCOVFA-AVGNSLFASA-N Leu-Leu-Glu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CCC(O)=O QNBVTHNJGCOVFA-AVGNSLFASA-N 0.000 description 1
- QMKFDEUJGYNFMC-AVGNSLFASA-N Leu-Pro-Arg Chemical compound CC(C)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCCN=C(N)N)C(O)=O QMKFDEUJGYNFMC-AVGNSLFASA-N 0.000 description 1
- BMVFXOQHDQZAQU-DCAQKATOSA-N Leu-Pro-Asp Chemical compound CC(C)C[C@@H](C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(=O)O)C(=O)O)N BMVFXOQHDQZAQU-DCAQKATOSA-N 0.000 description 1
- ILDSIMPXNFWKLH-KATARQTJSA-N Leu-Thr-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(O)=O ILDSIMPXNFWKLH-KATARQTJSA-N 0.000 description 1
- WGAZVKFCPHXZLO-SZMVWBNQSA-N Leu-Trp-Glu Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N WGAZVKFCPHXZLO-SZMVWBNQSA-N 0.000 description 1
- HOMFINRJHIIZNJ-HOCLYGCPSA-N Leu-Trp-Gly Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)NCC(O)=O HOMFINRJHIIZNJ-HOCLYGCPSA-N 0.000 description 1
- WUHBLPVELFTPQK-KKUMJFAQSA-N Leu-Tyr-Asn Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(N)=O)C(O)=O WUHBLPVELFTPQK-KKUMJFAQSA-N 0.000 description 1
- SXOFUVGLPHCPRQ-KKUMJFAQSA-N Leu-Tyr-Cys Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CS)C(O)=O SXOFUVGLPHCPRQ-KKUMJFAQSA-N 0.000 description 1
- DEFGUIIUYAUEDU-ZPFDUUQYSA-N Lys-Asn-Ile Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O DEFGUIIUYAUEDU-ZPFDUUQYSA-N 0.000 description 1
- DRRXXZBXDMLGFC-IHRRRGAJSA-N Lys-Val-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CCCCN DRRXXZBXDMLGFC-IHRRRGAJSA-N 0.000 description 1
- IHITVQKJXQQGLJ-LPEHRKFASA-N Met-Asn-Pro Chemical compound CSCC[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N1CCC[C@@H]1C(=O)O)N IHITVQKJXQQGLJ-LPEHRKFASA-N 0.000 description 1
- FJVJLMZUIGMFFU-BQBZGAKWSA-N Met-Asp-Gly Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)NCC(O)=O FJVJLMZUIGMFFU-BQBZGAKWSA-N 0.000 description 1
- RXWPLVRJQNWXRQ-IHRRRGAJSA-N Met-His-His Chemical compound C([C@H](NC(=O)[C@@H](N)CCSC)C(=O)N[C@@H](CC=1N=CNC=1)C(O)=O)C1=CNC=N1 RXWPLVRJQNWXRQ-IHRRRGAJSA-N 0.000 description 1
- GETCJHFFECHWHI-QXEWZRGKSA-N Met-Ile-Gly Chemical compound CC[C@H](C)[C@@H](C(=O)NCC(=O)O)NC(=O)[C@H](CCSC)N GETCJHFFECHWHI-QXEWZRGKSA-N 0.000 description 1
- TWEWRDAAIYBJTO-ULQDDVLXSA-N Met-Tyr-His Chemical compound CSCC[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H](CC2=CN=CN2)C(=O)O)N TWEWRDAAIYBJTO-ULQDDVLXSA-N 0.000 description 1
- WUGMRIBZSVSJNP-UHFFFAOYSA-N N-L-alanyl-L-tryptophan Natural products C1=CC=C2C(CC(NC(=O)C(N)C)C(O)=O)=CNC2=C1 WUGMRIBZSVSJNP-UHFFFAOYSA-N 0.000 description 1
- XMBSYZWANAQXEV-UHFFFAOYSA-N N-alpha-L-glutamyl-L-phenylalanine Natural products OC(=O)CCC(N)C(=O)NC(C(O)=O)CC1=CC=CC=C1 XMBSYZWANAQXEV-UHFFFAOYSA-N 0.000 description 1
- 108010066427 N-valyltryptophan Proteins 0.000 description 1
- MQWISMJKHOUEMW-ULQDDVLXSA-N Phe-Arg-His Chemical compound C([C@H](N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CC=1NC=NC=1)C(O)=O)C1=CC=CC=C1 MQWISMJKHOUEMW-ULQDDVLXSA-N 0.000 description 1
- JIYJYFIXQTYDNF-YDHLFZDLSA-N Phe-Asn-Val Chemical compound CC(C)[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CC1=CC=CC=C1)N JIYJYFIXQTYDNF-YDHLFZDLSA-N 0.000 description 1
- APJPXSFJBMMOLW-KBPBESRZSA-N Phe-Gly-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CC1=CC=CC=C1 APJPXSFJBMMOLW-KBPBESRZSA-N 0.000 description 1
- HBGFEEQFVBWYJQ-KBPBESRZSA-N Phe-Gly-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CC1=CC=CC=C1 HBGFEEQFVBWYJQ-KBPBESRZSA-N 0.000 description 1
- MJAYDXWQQUOURZ-JYJNAYRXSA-N Phe-Lys-Gln Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(N)=O)C(O)=O MJAYDXWQQUOURZ-JYJNAYRXSA-N 0.000 description 1
- WDOCBGZHAQQIBL-IHPCNDPISA-N Phe-Trp-Ser Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CO)C(O)=O)C1=CC=CC=C1 WDOCBGZHAQQIBL-IHPCNDPISA-N 0.000 description 1
- CMOIIANLNNYUTP-SRVKXCTJSA-N Pro-Gln-His Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CC2=CN=CN2)C(=O)O CMOIIANLNNYUTP-SRVKXCTJSA-N 0.000 description 1
- NXEYSLRNNPWCRN-SRVKXCTJSA-N Pro-Glu-Leu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O NXEYSLRNNPWCRN-SRVKXCTJSA-N 0.000 description 1
- LXVLKXPFIDDHJG-CIUDSAMLSA-N Pro-Glu-Ser Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(O)=O LXVLKXPFIDDHJG-CIUDSAMLSA-N 0.000 description 1
- DRKAXLDECUGLFE-ULQDDVLXSA-N Pro-Leu-Phe Chemical compound CC(C)C[C@H](NC(=O)[C@@H]1CCCN1)C(=O)N[C@@H](Cc1ccccc1)C(O)=O DRKAXLDECUGLFE-ULQDDVLXSA-N 0.000 description 1
- KDBHVPXBQADZKY-GUBZILKMSA-N Pro-Pro-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@@H]1CCCN1C(=O)[C@H]1NCCC1 KDBHVPXBQADZKY-GUBZILKMSA-N 0.000 description 1
- BGWKULMLUIUPKY-BQBZGAKWSA-N Pro-Ser-Gly Chemical compound OC(=O)CNC(=O)[C@H](CO)NC(=O)[C@@H]1CCCN1 BGWKULMLUIUPKY-BQBZGAKWSA-N 0.000 description 1
- SNSYSBUTTJBPDG-OKZBNKHCSA-N Pro-Trp-Pro Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CC2=CNC3=CC=CC=C32)C(=O)N4CCC[C@@H]4C(=O)O SNSYSBUTTJBPDG-OKZBNKHCSA-N 0.000 description 1
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 1
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 1
- 241000220317 Rosa Species 0.000 description 1
- OYEDZGNMSBZCIM-XGEHTFHBSA-N Ser-Arg-Thr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(O)=O OYEDZGNMSBZCIM-XGEHTFHBSA-N 0.000 description 1
- WKLJLEXEENIYQE-SRVKXCTJSA-N Ser-Cys-Tyr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CS)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O WKLJLEXEENIYQE-SRVKXCTJSA-N 0.000 description 1
- VMLONWHIORGALA-SRVKXCTJSA-N Ser-Leu-Leu Chemical compound CC(C)C[C@@H](C([O-])=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H]([NH3+])CO VMLONWHIORGALA-SRVKXCTJSA-N 0.000 description 1
- PPNPDKGQRFSCAC-CIUDSAMLSA-N Ser-Lys-Asp Chemical compound NCCCC[C@H](NC(=O)[C@@H](N)CO)C(=O)N[C@@H](CC(O)=O)C(O)=O PPNPDKGQRFSCAC-CIUDSAMLSA-N 0.000 description 1
- PPCZVWHJWJFTFN-ZLUOBGJFSA-N Ser-Ser-Asp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(O)=O PPCZVWHJWJFTFN-ZLUOBGJFSA-N 0.000 description 1
- GYDFRTRSSXOZCR-ACZMJKKPSA-N Ser-Ser-Glu Chemical compound OC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCC(O)=O GYDFRTRSSXOZCR-ACZMJKKPSA-N 0.000 description 1
- PLQWGQUNUPMNOD-KKUMJFAQSA-N Ser-Tyr-Leu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(C)C)C(O)=O PLQWGQUNUPMNOD-KKUMJFAQSA-N 0.000 description 1
- SIEBDTCABMZCLF-XGEHTFHBSA-N Ser-Val-Thr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O SIEBDTCABMZCLF-XGEHTFHBSA-N 0.000 description 1
- XDARBNMYXKUFOJ-GSSVUCPTSA-N Thr-Asp-Thr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O XDARBNMYXKUFOJ-GSSVUCPTSA-N 0.000 description 1
- ODXKUIGEPAGKKV-KATARQTJSA-N Thr-Leu-Cys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CS)C(=O)O)N)O ODXKUIGEPAGKKV-KATARQTJSA-N 0.000 description 1
- MUAFDCVOHYAFNG-RCWTZXSCSA-N Thr-Pro-Arg Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCCNC(N)=N)C(O)=O MUAFDCVOHYAFNG-RCWTZXSCSA-N 0.000 description 1
- XGFYGMKZKFRGAI-RCWTZXSCSA-N Thr-Val-Arg Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CCCN=C(N)N XGFYGMKZKFRGAI-RCWTZXSCSA-N 0.000 description 1
- KPMIQCXJDVKWKO-IFFSRLJSSA-N Thr-Val-Glu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O KPMIQCXJDVKWKO-IFFSRLJSSA-N 0.000 description 1
- BKVICMPZWRNWOC-RHYQMDGZSA-N Thr-Val-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)[C@@H](C)O BKVICMPZWRNWOC-RHYQMDGZSA-N 0.000 description 1
- LTLBNCDNXQCOLB-UBHSHLNASA-N Trp-Asp-Ser Chemical compound C1=CC=C2C(C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O)=CNC2=C1 LTLBNCDNXQCOLB-UBHSHLNASA-N 0.000 description 1
- PKUJMYZNJMRHEZ-XIRDDKMYSA-N Trp-Glu-Arg Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O PKUJMYZNJMRHEZ-XIRDDKMYSA-N 0.000 description 1
- YCQKQFKXBPJXRY-PMVMPFDFSA-N Trp-Tyr-Lys Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CC3=CC=C(C=C3)O)C(=O)N[C@@H](CCCCN)C(=O)O)N YCQKQFKXBPJXRY-PMVMPFDFSA-N 0.000 description 1
- NIHNMOSRSAYZIT-BPNCWPANSA-N Tyr-Ala-Arg Chemical compound NC(=N)NCCC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 NIHNMOSRSAYZIT-BPNCWPANSA-N 0.000 description 1
- IIJWXEUNETVJPV-IHRRRGAJSA-N Tyr-Arg-Ser Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CO)C(=O)O)N)O IIJWXEUNETVJPV-IHRRRGAJSA-N 0.000 description 1
- RUCNAYOMFXRIKJ-DCAQKATOSA-N Val-Ala-Lys Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CCCCN RUCNAYOMFXRIKJ-DCAQKATOSA-N 0.000 description 1
- GNWUWQAVVJQREM-NHCYSSNCSA-N Val-Asn-His Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)N GNWUWQAVVJQREM-NHCYSSNCSA-N 0.000 description 1
- COSLEEOIYRPTHD-YDHLFZDLSA-N Val-Asp-Tyr Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 COSLEEOIYRPTHD-YDHLFZDLSA-N 0.000 description 1
- CVIXTAITYJQMPE-LAEOZQHASA-N Val-Glu-Asn Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O CVIXTAITYJQMPE-LAEOZQHASA-N 0.000 description 1
- CPGJELLYDQEDRK-NAKRPEOUSA-N Val-Ile-Ala Chemical compound CC[C@H](C)[C@H](NC(=O)[C@@H](N)C(C)C)C(=O)N[C@@H](C)C(O)=O CPGJELLYDQEDRK-NAKRPEOUSA-N 0.000 description 1
- ZHQWPWQNVRCXAX-XQQFMLRXSA-N Val-Leu-Pro Chemical compound CC(C)C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](C(C)C)N ZHQWPWQNVRCXAX-XQQFMLRXSA-N 0.000 description 1
- XPKCFQZDQGVJCX-RHYQMDGZSA-N Val-Lys-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](C(C)C)N)O XPKCFQZDQGVJCX-RHYQMDGZSA-N 0.000 description 1
- VCIYTVOBLZHFSC-XHSDSOJGSA-N Val-Phe-Pro Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N2CCC[C@@H]2C(=O)O)N VCIYTVOBLZHFSC-XHSDSOJGSA-N 0.000 description 1
- GTACFKZDQFTVAI-STECZYCISA-N Val-Tyr-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)C(C)C)CC1=CC=C(O)C=C1 GTACFKZDQFTVAI-STECZYCISA-N 0.000 description 1
- ZHWZDZFWBXWPDW-GUBZILKMSA-N Val-Val-Cys Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CS)C(O)=O ZHWZDZFWBXWPDW-GUBZILKMSA-N 0.000 description 1
- VVIZITNVZUAEMI-DLOVCJGASA-N Val-Val-Gln Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CCC(N)=O VVIZITNVZUAEMI-DLOVCJGASA-N 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 238000001042 affinity chromatography Methods 0.000 description 1
- 108010047495 alanylglycine Proteins 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 108010013835 arginine glutamate Proteins 0.000 description 1
- 108010052670 arginyl-glutamyl-glutamic acid Proteins 0.000 description 1
- 108010077245 asparaginyl-proline Proteins 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 230000009920 chelation Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 239000003480 eluent Substances 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000000834 fixative Substances 0.000 description 1
- 238000002189 fluorescence spectrum Methods 0.000 description 1
- 238000001215 fluorescent labelling Methods 0.000 description 1
- 102000034287 fluorescent proteins Human genes 0.000 description 1
- 108091006047 fluorescent proteins Proteins 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- 108010013768 glutamyl-aspartyl-proline Proteins 0.000 description 1
- 108010079547 glutamylmethionine Proteins 0.000 description 1
- 108010020688 glycylhistidine Proteins 0.000 description 1
- 108010037850 glycylvaline Proteins 0.000 description 1
- 239000011544 gradient gel Substances 0.000 description 1
- 108010040030 histidinoalanine Proteins 0.000 description 1
- 108010036413 histidylglycine Proteins 0.000 description 1
- 108010092114 histidylphenylalanine Proteins 0.000 description 1
- 108010085325 histidylproline Proteins 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 125000004435 hydrogen atom Chemical group [H]* 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 108010044311 leucyl-glycyl-glycine Proteins 0.000 description 1
- 108010073472 leucyl-prolyl-proline Proteins 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000012160 loading buffer Substances 0.000 description 1
- 108010009298 lysylglutamic acid Proteins 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 108010063431 methionyl-aspartyl-glycine Proteins 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 230000005311 nuclear magnetism Effects 0.000 description 1
- 239000003208 petroleum Substances 0.000 description 1
- 108010012581 phenylalanylglutamate Proteins 0.000 description 1
- 230000004853 protein function Effects 0.000 description 1
- 238000001742 protein purification Methods 0.000 description 1
- 230000012743 protein tagging Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000010453 quartz Substances 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 229930187593 rose bengal Natural products 0.000 description 1
- 229940081623 rose bengal Drugs 0.000 description 1
- STRXNPAVPKGJQR-UHFFFAOYSA-N rose bengal A Natural products O1C(=O)C(C(=CC=C2Cl)Cl)=C2C21C1=CC(I)=C(O)C(I)=C1OC1=C(I)C(O)=C(I)C=C21 STRXNPAVPKGJQR-UHFFFAOYSA-N 0.000 description 1
- 108010069117 seryl-lysyl-aspartic acid Proteins 0.000 description 1
- 108010026333 seryl-proline Proteins 0.000 description 1
- 108010071207 serylmethionine Proteins 0.000 description 1
- 238000010898 silica gel chromatography Methods 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicon dioxide Inorganic materials O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical class O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 239000012086 standard solution Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 238000010189 synthetic method Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 229910052723 transition metal Inorganic materials 0.000 description 1
- 229910001428 transition metal ion Inorganic materials 0.000 description 1
- 108010035534 tyrosyl-leucyl-alanine Proteins 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- 238000000870 ultraviolet spectroscopy Methods 0.000 description 1
Images
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
Abstract
A histidine-tagged fluorescent probe, a preparation method and application thereof. The invention discloses a rhodamine B derivative-based histidine-tagged fluorescent compound, and a preparation method and application thereof. The fluorescent compound Rho-IDA can chelate different metal ions, wherein the fluorescent probe derivative Rho-IDA-CoII obtained by chelating bivalent cobalt ions has the function of specifically marking histidine-tag fusion protein. The fluorescent probe has the advantages of simple and convenient synthesis method, small molecular weight and excitation and emission wavelengths in a near infrared region. When the method is applied to biological imaging, the interference of fluorescent impurities in the background environment can be reduced; meanwhile, the histidine-tagged fusion protein in the gel can be rapidly and specifically labeled and stained, and a reliable research tool is provided for related biochemical research.
Description
Technical Field
The invention relates to the technical field of chemistry and biology, in particular to a rhodamine B derivative-based histidine-tagged fluorescent compound and a preparation method and application thereof.
Background
Biochemical markers of proteins may elucidate the function, location and dynamics of proteins. The recombinant protein fluorescent marker based on small molecules is used as a substitute scheme of a fluorescent protein fusion technology, has a special prospect, and cannot cause harmful interference on protein functions. The typical technology is based on fluorescence imaging of small molecule labels, and the technology is designed and synthesized into a fluorescent probe capable of combining with a specific short peptide, and the protein labeling of the small molecule fluorescent probe is realized by fusing the specific short peptide to a target protein. In recent decades, the research of labeling proteins based on small molecule probes has been greatly advanced. Among them, the metal chelate labeling technique of proteins has been attracting attention because of its simplicity of operation and high specificity. Due to His6-Ni2+The NTA system is widely applied to affinity chromatography protein purification, so that the system can specifically label histidine tag (His-tag) fusion protein by coupling fluorescent groups. Currently common are fluorescent probes based on single or multiple NTA (nitrilotriacetate) derivative coupling, although these probes overcome the histidine tag with Ni2+Weak binding force. However, due to the large molecular weight of the fluorescent probes, the defects of high application cost, complex synthetic method, slow protein labeling and the like exist, so that the fluorescent probes cannot be applied to most of the existing histidine-tag fusion protein libraries. Therefore, the design of the histidine-tag fusion protein fluorescent probe with high sensitivity, good specificity and wide application range has important significance.
Disclosure of Invention
The invention aims to provide a rhodamine B derivative-based histidine-tagged fluorescent probe and a preparation method and application thereof.
The technical scheme adopted by the invention is as follows:
a fluorescent compound Rho-IDA based on rhodamine B derivatives has a structural formula shown as (I):
the invention also relates to a method for preparing the fluorescent compound Rho-IDA, which comprises the following steps: reacting rhodamine B with N-hydroxysuccinimide to generate rhodamine B-NHS ester, and reacting the rhodamine B-NHS ester with iminodiacetic acid to generate Rho-IDA.
Specifically, the method comprises the following steps:
(1) mixing rhodamine B and N-hydroxysuccinimide, reacting overnight at room temperature, fully activating carboxyl of rhodamine B, and filtering to remove impurities;
(2) adding iminodiacetic acid and triethylamine into the reaction liquid obtained in the step (1), and reacting at room temperature overnight;
(3) adjusting the pH value of the reaction solution to 2.0 by using HCl solution, and stirring at room temperature until a precipitate is separated out;
(4) and filtering, collecting precipitates, rinsing the precipitates by using an HCl solution, and evaporating the solvent to obtain the fluorescent compound Rho-IDA.
The invention also relates to a metal chelate Rho-IDA-M of the fluorescent compound Rho-IDA, which has a structural formula shown as (II):
in formula (II), M is a metal element capable of being chelated by the fluorescent probe Rho-IDA, such as a transition metal element.
The metal chelate Rho-IDA-M is formed by the equimolar M of a fluorescent compound Rho-IDA2+Mixing uniformly and carrying out chelation reaction.
Preferably, M is one of the following: cu2+，Co2+，Ni2+。
The invention also relates to the application of the fluorescent compound Rho-IDA and the metal chelate Rho-IDA-M thereof in the preparation of fluorescent probes.
Specifically, the fluorescent probe is used for specifically and fluorescently labeling the histidine-tag fusion protein, or is used for quantitatively detecting the trace histidine-tag fusion protein.
The invention has the following beneficial effects: the invention designs and synthesizes a novel histidine-tagged fluorescent compound based on IDA (iminodiacetic acid) coupled rhodamine B, and the fluorescent compound as a fluorescent probe has the advantages that the excitation wavelength and the emission wavelength are positioned in a near infrared region, the interference of the fluorescence background of a sample can be reduced when the fluorescent probe is applied to fluorescence imaging, the sensitivity is high, the specificity is strong, and 1 pmol of histidine-tagged protein can be specifically detected; the fluorescent probe can be rapidly prepared by a two-step method under the conditions of normal temperature and normal pressure, can be combined with different divalent transition metal ions, and provides a reliable research tool in the research of the interaction of the fluorescence labeling metal ions.
Drawings
FIG. 1 is a nuclear magnetic hydrogen spectrum of the probe Rho-IDA of the present invention.
FIG. 2 is an ESI-MS spectrum of a probe Rho-IDA of the present invention.
FIG. 3 is a fluorescence spectrum of the probe Rho-IDA-CoII of the present invention.
Detailed Description
The invention will be further described with reference to specific examples, but the scope of the invention is not limited thereto:
test materials and reagents:
1. the analytical purification chemical reagents used in the present invention were purchased from chemical reagents of the national pharmaceutical group, ltd.
2. SDS-PAGE Experimental instruments and reagents: SurePAGETMThe gradient concentration (4-20%) prefabricated glue is purchased from Nanjing Kingsrie Biotech, Inc.; the constant voltage electrophoresis uses Berkeli-Protean®A Tetra electrophoresis tank; gel imaging Using Burle ChemiDoc XRS+Provided is a system.
3. Fixing agent: deionized water, ethanol and acetic acid are prepared according to the proportion of 10:5:1 and are fully and uniformly mixed.
4. Fluorescent dye solution: Rho-IDA-CoII 1. mu.M, Tris-HCl buffer (pH 8.0) 10 mM and NaCl 300 mM.
5. Washing liquid: imidazole 50 mM, Tris-HCl buffer (pH 8.0) 10 mM and NaCl 300 mM.
Description of the drawings: the molecular biological experiments, which are not specifically described in the following examples, were performed by referring to the specific methods listed in molecular cloning, A laboratory Manual (third edition) J. SammBruk, or according to the kit and product instructions
Example 1: chemical synthesis of fluorescent probe Rho-IDA
1) In a 50 mL reaction flask equipped with a magnetic stirrer, 0.1 mmol of rhodamine B and 10 mL of dichloromethane as raw materials were added and sufficiently dissolved.
2) Slowly adding 0.15 mmol of N-hydroxysuccinimide and 0.15 mmol of N, N' -dicyclohexylcarbodiimide, reacting at room temperature overnight, and filtering to remove precipitate.
3) And slowly adding 0.2 mmol of triethylamine, and uniformly mixing.
4) 0.15 mmol of iminodiacetic acid was added and the reaction was allowed to proceed overnight at room temperature.
5) The solvent was evaporated under reduced pressure, and the residue was dissolved in 30 mL of methylene chloride, followed by addition of 30 mL of saturated brine, extraction and separation of the methylene chloride layer.
6) Filtering to remove impurities, and performing silica gel column chromatography with petroleum ether and ethyl acetate =5:1 as eluent to obtain rose bengal powder with a total yield of 64%.
The synthetic products obtained above were identified using a Bruker Avance III spectrometer and a Therm LCQ TM Deca XPplus mass spectrometer, the results of which are shown in FIGS. 1 and 2. The identification result of the electrospray ionization mass spectrum (ESI-MS) is as follows: m/z: 558.46, calculated value of Rho-IDA [ M]+: 558, match; nuclear magnetism of1H-NMR identifies that the number of hydrogen atoms is consistent with the structure of the fluorescent probe. The result shows that the rose red powder synthesized by the method is the fluorescent probe Rho-IDA.
Example 2: fluorescence property determination of fluorescent probe Rho-IDA-CoII
1) Rho-IDA and CoCl at the same molar concentration2Equivalently and uniformly mixing to obtain a fluorescent probe Rho-IDA-CoII;
2) and (3) taking 200 mu L of 0.1 mM fluorescent probe Rho-IDA-CoII aqueous solution into a trace quartz cuvette, and performing full-wavelength scanning on the Rho-IDA-CoII sample by using an ultraviolet-visible spectrophotometer, wherein the wavelength range is 200 nm-850 nm, and ultrapure water is used as a blank control. The full-wavelength scanning result shows that the fluorescent probe Rho-IDA-CoII has maximum absorption peaks at two wavelengths of 550 nm and 575 nm.
3) Transferring the fluorescent probe Rho-IDA-CoII solution sample in the step 2 into a micro-fluorescent cuvette, setting the excitation wavelength to be 550 nm, and scanning the fluorescence intensity of the fluorescent probe Rho-IDA-CoII within the wavelength range of 400-750 nm;
4) the detection result shows that (figure 3) the fluorescent probe Rho-IDA-CoII can emit strong fluorescence at 580 nm under the excitation of visible light with the wavelength of 550 nm.
Example 3: histidine-tagged protein in vitro staining based on fluorescent probe Rho-IDA-CoII
1) Uniformly mixing histidine tag fusion protein (SEQ ID number 1) to be detected and electrophoresis loading buffer solution in a ratio of 3:1, and carrying out boiling water bath for 5 min;
2) performing SDS-PAGE separation on the protein sample by using 4-20% gradient gel (Kinry, China), wherein the electrophoresis condition is constant voltage of 200V and 40 min;
3) after electrophoresis is finished, carefully placing the gel in a fixing solution to be soaked for 1 h;
4) rinsing the gel with deionized water for 2 times, each time for 10 min, and sufficiently removing the fixative;
5) transferring the gel into a fluorescent dye solution to dye for 1 h in a dark place;
6) rinsing the gel with washing solution for 10 min for 2 times;
7) capturing a fluorescent gel image using a berle CCD camera;
8) dyeing the gel with the recorded fluorescent dyeing result in the step 7 by using Coomassie brilliant blue, and comparing and analyzing the fluorescent dyeing effect;
9) the result shows that the fluorescent probe Rho-IDA-CoII can specifically identify and mark the histidine-tagged fusion protein. Compared with a Coomassie brilliant blue staining result, the marking staining of the fluorescent probe Rho-IDA-CoII on the histidine-tagged fusion protein is not substantially interfered by non-tagged protein, and trace histidine-tagged fusion protein can be clearly marked.
Example 4: fluorescent probe Rho-IDA-CoII detection sensitivity determination
1) Accurately preparing standard solutions of histidine-tagged fusion protein (SEQ ID number 1) with the concentrations of 1, 2, 3, 5, 10, 50, 100, 150 and 200 ng/. mu.L respectively;
2) SDS-PAGE analysis of the BSA sample was performed according to the method of example 3, and the loading of the standard protein sample was 10. mu.L;
3) after a fluorescent probe Rho-IDA-CoII is used for marking, the fluorescent staining effect is detected;
4) gel imaging results show that when the protein loading is above 30 ng (about 1 pmol), a band clearly imaged by the instrument is emitted. The detection sensitivity of the fluorescent probe Rho-IDA-CoII is 1 pmol histidine-tagged fusion protein.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Sequence listing
<110> Zhejiang industrial university
<120> histidine tag fluorescent probe and preparation method and application thereof
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 443
<212> PRT
<213> Unknown (Unknown)
<400> 1
Met His His His His His His His His His His Asn Gly Ile His Asp
1 5 10 15
Leu Gly Gly Met Asp Gly Phe Gly Lys Ile Ile Arg Glu Glu Asn Glu
20 25 30
Pro Leu Phe His Lys Asp Trp Glu Arg Ile Ala Phe Gly Leu Leu Ile
35 40 45
Gly Thr Ala Gly Gln Gly Leu Tyr Asn Leu Asp Glu Phe Arg His Ala
50 55 60
Ile Glu Arg Met Asn Pro Val Asp Tyr Leu Thr Ser Gly Tyr Tyr Gly
65 70 75 80
His Trp Val Ala Ser Ile Ala Thr Leu Leu Val Glu Lys Gly Ile Leu
85 90 95
Asp Ala Ser Glu Leu Val Ser Arg Thr Gln Thr Tyr Leu Ala Gln Pro
100 105 110
Asp Thr Lys Thr Pro Arg Arg Glu Asn Pro Glu Leu Val Asn His Leu
115 120 125
Glu Gln Val Ile Lys Val Gly Val Ser Thr Val Arg Glu Val Ser Ser
130 135 140
Ala Pro Arg Phe Asn Val Gly Asp Arg Val Lys Thr Lys Asn Ile His
145 150 155 160
Pro Ser Gly His Thr Arg Leu Pro Arg Tyr Ala Arg Asp Lys Tyr Gly
165 170 175
Val Ile Ala Met Tyr His Gly Ala His Val Phe Pro Asp Ala Asn Ala
180 185 190
His Gly Lys Gly Glu Ser Pro Gln His Leu Tyr Cys Ile Arg Phe Glu
195 200 205
Ala Asn Glu Leu Trp Gly Ile Gln Gln Gly Glu Ala Val Tyr Ile Asp
210 215 220
Leu Trp Glu Ser Tyr Leu Glu Pro Val Ser Lys Asp Asn Asn Lys Val
225 230 235 240
His His His His Pro His Pro Glu Ser Phe Trp Ser Ala Arg Ala Lys
245 250 255
Ala Leu Glu Ser Leu Leu Ile Glu Lys Gly Ile Leu Ser Ser Asp Ala
260 265 270
Ile Asp Arg Val Val Gln His Tyr Glu His Glu Leu Gly Pro Met Asn
275 280 285
Gly Ala Lys Val Val Ala Lys Ala Trp Thr Asp Pro Ala Phe Lys Gln
290 295 300
Arg Leu Leu Glu Asp Pro Glu Thr Val Leu Arg Glu Leu Gly Tyr Tyr
305 310 315 320
Gly Leu Gln Gly Glu His Ile Arg Val Val Glu Asn Thr Asp Thr Val
325 330 335
His Asn Val Val Val Cys Thr Leu Cys Ser Cys Tyr Pro Trp Pro Leu
340 345 350
Leu Gly Leu Pro Pro Ala Trp Tyr Lys Glu Pro Thr Tyr Arg Ser Arg
355 360 365
Ile Val Lys Glu Pro Arg Lys Val Leu Arg Glu Glu Phe Gly Leu Asp
370 375 380
Leu Pro Asp Thr Val Glu Ile Arg Val Trp Asp Ser Ser Ser Glu Met
385 390 395 400
Arg Tyr Met Val Leu Pro Gln Arg Pro Glu Gly Thr Glu Gly Met Thr
405 410 415
Glu Glu Glu Leu Ala Lys Ile Val Thr Arg Asp Ser Met Ile Gly Val
420 425 430
Ala Lys Val Gln Pro Ser Ser Val Thr Val Arg
435 440
Claims (8)
1. A fluorescent compound Rho-IDA based on rhodamine B derivatives has a structural formula shown as (I):
2. a method for preparing the fluorescent compound Rho-IDA of claim 1, wherein the method comprises: reacting rhodamine B with N-hydroxysuccinimide to generate rhodamine B-NHS ester, and reacting the rhodamine B-NHS ester with iminodiacetic acid to generate Rho-IDA.
3. The method of claim 2, wherein the method comprises:
(1) mixing rhodamine B and N-hydroxysuccinimide, reacting overnight at room temperature, and filtering to remove impurities;
(2) adding iminodiacetic acid and triethylamine into the reaction liquid obtained in the step (1), and reacting at room temperature overnight;
(3) adjusting the pH value of the reaction solution to 2.0 by using HCl solution, and stirring at room temperature until a precipitate is separated out;
and filtering, collecting precipitates, rinsing the precipitates by using an HCl solution, and evaporating the solvent to obtain the fluorescent compound Rho-IDA.
4. A metal chelate Rho-IDA-M of a fluorescent compound Rho-IDA, which has a structural formula shown as (II): (II) formula
In (II), M is a metal element capable of being chelated by the fluorescent compound Rho-IDA.
5. A metal chelate Rho-IDA-M according to claim 3, characterized in that said M is one of the following: cu2+，Co2+，Ni2+。
6. The application of a fluorescent compound Rho-IDA and a metal chelate Rho-IDA-M thereof in preparing a fluorescent probe.
7. The use according to claim 5, wherein the fluorescent probe is used to specifically fluorescently label a histidine-tag fusion protein.
8. The use according to claim 5, wherein the fluorescent probe is used for quantitative detection of a trace amount of histidine-tagged fusion protein.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111440062.0A CN114315784A (en) | 2021-11-30 | 2021-11-30 | Histidine-labeled fluorescent probe and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111440062.0A CN114315784A (en) | 2021-11-30 | 2021-11-30 | Histidine-labeled fluorescent probe and preparation method and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN114315784A true CN114315784A (en) | 2022-04-12 |
Family
ID=81047996
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111440062.0A Pending CN114315784A (en) | 2021-11-30 | 2021-11-30 | Histidine-labeled fluorescent probe and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114315784A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116990512A (en) * | 2023-09-25 | 2023-11-03 | 聚诚(北京)生物科技有限责任公司 | Matrix metalloproteinase-9 detection kit |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2008145609A1 (en) * | 2007-05-25 | 2008-12-04 | Novartis Ag | Method of making covalent conjugates with his-tagged proteins |
| CN106905418A (en) * | 2017-01-18 | 2017-06-30 | 华东理工大学 | A kind of histidine fluorescence probe and its preparation method and application |
| CN109810138A (en) * | 2018-12-26 | 2019-05-28 | 浙江工业大学 | A kind of targetted mitochondria Small-molecule probe and its preparation method and application |
| CN110128394A (en) * | 2019-03-25 | 2019-08-16 | 天津农学院 | A kind of fluorescent chemicals and its application for the detection of underwater gold category ion concentration |
| CN111533724A (en) * | 2020-05-28 | 2020-08-14 | 浙江昂拓莱司生物技术有限公司 | His-tag fluorescent probe with near-infrared property and preparation and application thereof |
-
2021
- 2021-11-30 CN CN202111440062.0A patent/CN114315784A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2008145609A1 (en) * | 2007-05-25 | 2008-12-04 | Novartis Ag | Method of making covalent conjugates with his-tagged proteins |
| CN106905418A (en) * | 2017-01-18 | 2017-06-30 | 华东理工大学 | A kind of histidine fluorescence probe and its preparation method and application |
| CN109810138A (en) * | 2018-12-26 | 2019-05-28 | 浙江工业大学 | A kind of targetted mitochondria Small-molecule probe and its preparation method and application |
| CN110128394A (en) * | 2019-03-25 | 2019-08-16 | 天津农学院 | A kind of fluorescent chemicals and its application for the detection of underwater gold category ion concentration |
| CN111533724A (en) * | 2020-05-28 | 2020-08-14 | 浙江昂拓莱司生物技术有限公司 | His-tag fluorescent probe with near-infrared property and preparation and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| 陈磊等: "特异性的蛋白小分子荧光探针及其标记技术", 《生命科学》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116990512A (en) * | 2023-09-25 | 2023-11-03 | 聚诚(北京)生物科技有限责任公司 | Matrix metalloproteinase-9 detection kit |
| CN116990512B (en) * | 2023-09-25 | 2023-12-08 | 聚诚(北京)生物科技有限责任公司 | Matrix metalloproteinase-9 detection kit |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP3371194B1 (en) | Protein sequencing method | |
| Yang et al. | Thiol–chromene click chemistry: A coumarin-based derivative and its use as regenerable thiol probe and in bioimaging applications | |
| Clede et al. | Detection of an estrogen derivative in two breast cancer cell lines using a single core multimodal probe for imaging (SCoMPI) imaged by a panel of luminescent and vibrational techniques | |
| Liu et al. | A novel near-infrared fluorescent probe with a large Stokes shift for biothiol detection and application in in vitro and in vivo fluorescence imaging | |
| Huo et al. | A turn-on green fluorescent thiol probe based on the 1, 2-addition reaction and its application for bioimaging | |
| Zhang et al. | Photoluminescent and electrochemiluminescent dual-signaling probe for bio-thiols based on a ruthenium (II) complex | |
| CN110092773B (en) | Xanthene derivative and preparation method and application thereof | |
| EP3160971B1 (en) | Method for selective detection and estimation of histidine and cystein | |
| CN109400609B (en) | Protein-labeled fluorescent probe for marking SNAP-tag | |
| CN106929006B (en) | It is a kind of using naphthalimide as the identification cysteine of parent nucleus and homocysteine fluorescence probe and its preparation and application | |
| Chen et al. | The fluorescent biomarkers for lipid droplets with quinolone-coumarin unit | |
| Chao et al. | A Ni-NTA-based red fluorescence probe for protein labelling in live cells | |
| Lu et al. | Ultrafast deep-red emission fluorescent probe for highly selective imaging of endogenous cysteine in living cells and mice | |
| Wang et al. | A novel reaction-based fluorescent turn-on probe for biothiols and its application in cell imaging | |
| US20240201196A1 (en) | Non-covalent halotag ligands | |
| CN114315784A (en) | Histidine-labeled fluorescent probe and preparation method and application thereof | |
| Fedorowicz et al. | Synthesis and evaluation of dihydro-[1, 2, 4] triazolo [4, 3-a] pyridin-2-ium carboxylates as fixed charge fluorescent derivatization reagents for MEKC and MS proteomic analyses | |
| Wang | Molecular engineering of an efficient iminocoumarin-based probe for practical sensing applications | |
| CN114605343A (en) | Fluorescent group LAN-OH, fluorescent sensor LAN-beta gal, preparation method and application thereof | |
| CN111362958B (en) | Rhodamine fluorescent probe for specifically recognizing reduced GSH (glutathione), and preparation method and application thereof | |
| WO2006132030A1 (en) | Novel compound, reagent comprising the compound for peptide or protein analysis, and method of analysis with the analytical reagent | |
| CN109206351A (en) | A kind of near infrared fluorescent probe, preparation method and application for surveying palladium ion based on flower cyanines structure | |
| CN110186891B (en) | Polypeptide fluorescent probe specifically combining copper ions and cysteine | |
| CN108530457A (en) | Amino acid detection reagent naphthalimide derivative and its synthetic method and application | |
| Rong et al. | A naphthalimide-indole fused chromophore-based fluorescent probe for the detection of biothiol with red emission and a large Stokes shift |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20220412 |
|
| RJ01 | Rejection of invention patent application after publication |